Mannitol and the Combination of Mannitol and Gelatin Impair Whole Blood Coagulation and the Platelet Function In Vitro

Abstract Background The mechanisms of trauma induced coagulopathy (TIC) are considered multifactorial. Amongst others, however, shedding of the endothelial glycocalyx resulting in increased concentrations of glycocalyx fragments in plasma might also play a role. Thus, we hypothesized that shedded glycocalyx components affect coagulation and may act as humoral mediators of TIC. Methods To investigate effects of heparan sulfate, chondroitin sulfate, syndecan-1, versican, and thrombomodulin we added these fragments to in vitro assays of whole blood from healthy volunteers to yield concentrations observed in trauma patients. Platelet function, whole blood coagulation, and fibrinolysis were measured by standard coagulation tests, impedance aggregometry (IA), and viscoelastic tests (VET). To assess dose-response relationships, we performed IA with increasing concentrations of versican and VET with increasing concentrations of thrombomodulin. Results Intrinsically activated clotting times (i.e., activated partial thromboplastin time and intrinsically activated VET with and without heparinase) were unaffected by any glycocalyx fragment. Thrombomodulin, however, significantly and dose-dependently diminished fibrinolysis as assessed by VET with exogenously added rt-PA, and increased rt-PA-induced lysis Indices after 30 (up to 108% of control, p < 0,0001), 45 (up to 368% of control, p < 0,0001), and 60 min (up to 950% of control, p < 0,0001) in VET. Versican impaired platelet aggregation in response to arachidonic acid (up to − 37,6%, p < 0,0001), ADP (up to − 14,5%, p < 0,0001), and collagen (up to − 31,8%, p < 0,0001) in a dose-dependent manner, but did not affect TRAP-6 induced platelet aggregation. Clotting time in extrinsically activated VET was shortened by heparan sulfate (− 7,2%, p = 0,024), chondroitin sulfate (− 11,6%, p = 0,016), versican (− 13%, p = 0,012%), and when combined (− 7,2%, p = 0,007). Conclusions Glycocalyx components exert distinct inhibitory effects on platelet function, coagulation, and fibrinolysis. These data do not support a ‘heparin-like auto-anticoagulation’ by shed glycosaminoglycans but suggest a possible role of versican in trauma-induced thrombocytopathy and of thrombomodulin in trauma-associated impairment of endogenous fibrinolysis.

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In vitro effects of 3 % hypertonic saline and 20 % mannitol on canine whole blood coagulation and platelet function

BMC Veterinary Research ◽

10.1186/s12917-015-0555-x ◽

2015 ◽

Vol 11 (1) ◽

Cited By ~ 12

Author(s):

Katja-Nicole Adamik ◽

Emmanuelle Butty ◽

Judith Howard

Keyword(s):

Blood Coagulation ◽

Hypertonic Saline ◽

Platelet Function ◽

Whole Blood ◽

In Vitro Effects

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Effect of Aspirin Infusions on Platelet Function in Humans

Clinical Science ◽

10.1042/cs0790037 ◽

1990 ◽

Vol 79 (1) ◽

pp. 37-42 ◽

Cited By ~ 8

Author(s):

K. M. Wilson ◽

D. M. Siebert ◽

E. M. Duncan ◽

A. A. Somogyi ◽

J. V. Lloyd ◽

...

Keyword(s):

Platelet Aggregation ◽

Blood Coagulation ◽

Platelet Function ◽

Whole Blood ◽

Dependent Manner ◽

Inhibitory Effects ◽

Concentration Dependent Manner ◽

Infusion Period

1. The inhibitory effects of aspirin on platelet function in vitro have been shown to be both time (over 3 h) and concentration (1–10 μmol/l) dependent. 2. To determine if these effects occurred in vivo, four volunteers received intravenous infusions on four occasions, to give constant plasma aspirin concentrations of 0, 1, 2 and 4 μmol/l over 3 h. Infusions were performed at intervals of at least 2 weeks. 3. Before and during the infusions, blood was taken for assay of aspirin concentrations, and measurements of platelet aggregation in response to collagen, adenosine 5′-pyrophosphate and arachidonate. Thromboxane generation after stimulated platelet aggregation and whole-blood coagulation was also measured. 4. At each aspirin concentration, both platelet aggregation and thromboxane generation in response to collagen and arachidonate were inhibited progressively over the 3 h infusion period. Greatest inhibition was seen during the 4 μmol/l infusion, which produced maximal or near-maximal inhibition by the third hour. 5. Thromboxane generated during whole-blood coagulation was similarly inhibited in both a time- and concentration-dependent manner throughout all aspirin infusions. 6. The progressive nature of the inhibition of platelet function with these low aspirin concentrations may be due to either slow aspirin transport across the platelet membrane or delayed interaction with cyclo-oxygenase.

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Comparison of the in vitro effects of saline, hypertonic hydroxyethyl starch, hypertonic saline, and two forms of hydroxyethyl starch on whole blood coagulation and platelet function in dogs

Journal of Veterinary Emergency and Critical Care ◽

10.1111/vec.12320 ◽

2015 ◽

Vol 25 (4) ◽

pp. 474-487 ◽

Cited By ~ 23

Author(s):

Virginie A. Wurlod ◽

Judith Howard ◽

Thierry Francey ◽

Ariane Schweighauser ◽

Katja N. Adamik

Keyword(s):

Blood Coagulation ◽

Hypertonic Saline ◽

Hydroxyethyl Starch ◽

Platelet Function ◽

Whole Blood ◽

In Vitro Effects

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Results of in vitro whole blood coagulation assays using ROTEM and the flow-chamber T-TAS system are affected by hematocrit

Thrombosis Research ◽

10.1016/j.thromres.2020.06.030 ◽

2020 ◽

Vol 194 ◽

pp. 98-100

Author(s):

Anna Ågren ◽

Gustaf Edgren ◽

Paul Hjemdahl ◽

Gunilla Gryfelt ◽

Anders Östlund ◽

...

Keyword(s):

Blood Coagulation ◽

Whole Blood ◽

Flow Chamber ◽

Coagulation Assays

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Thrombin generation and fibrin clot formation under hypothermic conditions: An in vitro evaluation of tissue factor initiated whole blood coagulation

Journal of Critical Care ◽

10.1016/j.jcrc.2013.10.010 ◽

2014 ◽

Vol 29 (1) ◽

pp. 24-30 ◽

Cited By ~ 9

Author(s):

Matthew F. Whelihan ◽

Armin Kiankhooy ◽

Kathleen E. Brummel-Ziedins

Keyword(s):

Tissue Factor ◽

Blood Coagulation ◽

Whole Blood ◽

Thrombin Generation ◽

Fibrin Clot ◽

Clot Formation ◽

In Vitro Evaluation

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Sevoflurane Inhibits Unstimulated and Agonist-induced Platelet Antigen Expression and Platelet Function in Whole Blood In Vitro

Anesthesiology ◽

10.1097/00000542-200111000-00028 ◽

2001 ◽

Vol 95 (5) ◽

pp. 1220-1225 ◽

Cited By ~ 9

Author(s):

Nicola A. Horn ◽

Lothar de Rossi ◽

Tilo Robitzsch ◽

Klaus E. Hecker ◽

Gabriele Hutschenreuter ◽

...

Keyword(s):

Platelet Function ◽

Whole Blood ◽

Control Sample ◽

Antigen Expression ◽

Adenosine Diphosphate ◽

Color Flow ◽

Platelet Antigen ◽

Function Analyzer ◽

Platelet Function Analyzer

Background Previous studies have reported conflicting results about the effect of sevoflurane on platelet aggregation. To clarify this point, we investigated the effects of sevoflurane on platelet antigen expression and function in vitro. Methods Human whole blood was incubated for 1 h with 0.5 and 1 minimum alveolar concentration sevoflurane, 21% O(2), and 5% CO(2). A control sample was kept at the same conditions without sevoflurane. After stimulation with adenosine diphosphate or thrombin receptor agonist peptide 6, samples were stained with fluorochrome conjugated antibodies, and the expression of platelet glycoproteins GPIIb/IIIa, GPIb, and P-selectin, as well as activated GPIIb/IIIa, were measured with two-color flow cytometry. In addition, platelet function was assessed by means of thromboelastography and using the platelet function analyzer 100. Results Already in subanesthetic concentrations, sevoflurane inhibits unstimulated and agonist-induced GPIIb/IIIa surface expression and activated GPIIb/IIIa expression on platelets in whole blood. The agonist-induced redistribution of GPIb into the open canalicular system was also impaired by sevoflurane, whereas no effect on P-selectin expression in activated platelets could be found. Sevoflurane significantly reduced the maximum thromboelastographic amplitude. Furthermore, platelet function analyzer 100 closure times were significantly prolonged. Conclusion The results show that sevoflurane significantly impairs platelet antigen expression in vitro. It is especially the inhibition of GPIIb/IIIa expression and activation that impairs bleeding time as reflected in thromboelastographic measurements and platelet function analyzer 100 closure times. The exact inhibitory mechanism remains unclear.

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ABO Mismatched Platelet, Plasma and Cryoprecipitate Transfusions and Bleeding in Transfused Surgical Patients.

Blood ◽

10.1182/blood.v106.11.4166.4166 ◽

2005 ◽

Vol 106 (11) ◽

pp. 4166-4166

Author(s):

Neil Blumberg ◽

Kelly F. Gettings ◽

Joanna M. Heal

Keyword(s):

Platelet Function ◽

Whole Blood ◽

Blood Platelets ◽

Blood Groups ◽

Surgical Patients ◽

Red Cell ◽

Group A ◽

Red Cell Transfusion ◽

Platelet Transfusions

Abstract Three observations led us to investigate whether infusion of ABO non-identical platelets might impair, rather than improve hemostasis. (1) Exposure of platelets to immune complexes or platelet specific antibody can interfere with platelet function in vitro (Thromb Haemost76: 774, 1996). (2) In surgical patients receiving similar numbers of platelet transfusions, those receiving ABO non-identical platelets require 50% more red cell transfusions (Transfusion41: 790, 2001). (3) Patients with acute leukemia receiving prophylactic platelet transfusions typically are reported with serious bleeding at a rate of 15–20%, yet the bleeding rate in patients receiving only ABO identical platelets is below 5% (BMC Blood Disorders4: 6, 2004). In this study, the number of red cell transfusions and clinical outcomes during March 2002-Feb 2003 for all surgical patients of blood groups B and AB (B/AB) who received platelet transfusions were compared with patients of blood groups O and A (O/A). Recipients of blood groups B/AB would not be expected to experience excess bleeding, as measured by red cell transfusions, compared with patients of blood groups O/A. However, because of the lower prevalence of blood groups B/AB in the donor blood supply B/AB recipients may more frequently receive ABO mismatched platelet transfusions. O/A surgical patients (n=281) who received platelet transfusions required a mean of 13 ± 13 (SD) red cell transfusions as compared with B/AB patients (n=54), who required 19 ± 25 red cells (p =0.0086). O/A patients also had shorter length of stay, mean = 25 ± 34 days as compared with B/AB patients at 36 ± 59 days (p =0.064). Rates of mortality and nosocomial infections were not statistically significantly different. O/A patients received a mean of 14 ± 19 units of whole blood platelets during and after surgery, compared with 16 ± 16 units for B/AB patients (p =0.47). O/A patients received a mean of 3.3 ± 6.2 ABO non-identical platelets in contrast with B/AB patients who received 7.5 ± 11 (p = 0.0001). Both groups received similar numbers of ABO identical platelets: 11 ± 16 (O/A) versus 9 ± 12 (B/AB) (p =0.35). All but two patients received only ABO identical FFP and both groups received similar total amounts of FFP (mean of 9 units versus 11 units). While O/A patients received similar mean amounts of cryoprecipitate (6 units) to B/AB patients (8 units), the B/AB patients received significantly more ABO non-identical cryoprecipitate (mean = 4.2 vs. 2.3 units; p = 0.02). To study the effects of ABO incompatible plasma on platelet function, we measured PFA-100 (epinephrine cartridge) closure times in reconstituted whole blood exposing group A platelets to either group A or O plasma. In four of seven instances, closure times for A platelets exposed to O plasma were prolonged by more than 50 seconds, compared with A platelets exposed to allogeneic A plasma. These preliminary results support previous observations that exposure to ABO non-identical platelet transfusions is associated with increased red cell transfusions. One possible mechanism is impaired platelet function caused by antibody or immune complex binding. We speculate that transfusion of ABO mismatched platelets, FFP and/or cryoprecipitate may in some instances exacerbate bleeding, rather than correcting defects in hemostasis. Though further investigation is needed before suggesting changes in clinical practice these findings raise the possibility that use of ABO identical blood components might reduce red cell transfusion needs in bleeding surgical patients.

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Predictive Value of Whole Blood and Plasma Coagulation Tests for Intra- and Postoperative Bleeding Risk: A Systematic Review

Seminars in Thrombosis and Hemostasis ◽

10.1055/s-0037-1602665 ◽

2017 ◽

Vol 43 (07) ◽

pp. 772-805 ◽

Cited By ~ 14

Author(s):

Julie Larsen ◽

Anne-Mette Hvas

Keyword(s):

Systematic Review ◽

Blood Coagulation ◽

Platelet Function ◽

Whole Blood ◽

Predictive Value ◽

Platelet Function Tests ◽

Predictive Values ◽

Function Tests ◽

Perioperative Bleeding ◽

Coagulation Tests

AbstractExcessive perioperative bleeding is associated with increased morbidity and mortality as well as increased economic costs. A range of whole blood laboratory tests for hemostatic monitoring has emerged, but their ability to predict perioperative bleeding is still debated. We conducted a systematic review of the existing literature assessing the ability of whole blood coagulation (thromboelastography [TEG]/thromboelastometry [ROTEM]/Sonoclot), platelet function tests, and standard plasma-based coagulation tests to predict bleeding in the perioperative setting. We searched PubMed and Embase, covering the period from 1966 to November 2016. In total, 99 original studies were included. The included studies assessed TEG/ROTEM/Sonoclot (n = 29), platelet function tests (n = 27), both test types (n = 8), and standard coagulation tests only (n = 18), and some (n = 17) investigated the predictive value of testing in patients receiving antithrombotic medication. In general, studies reported low positive predictive values for perioperative testing, whereas negative predictive values were high. The studies yielded moderate areas under receiver operator characteristics (ROC) curve (for the majority, 0.60–0.80). In conclusion, while useful in the diagnosis and management of patients with overt bleeding, whole blood coagulation and platelet function tests as well as standard coagulation tests demonstrated limited ability to predict perioperative bleeding in unselected patients. Therefore, we recommend that both whole blood and plasma-based coagulation tests are primarily used in case of bleeding and not for screening in unselected patients prior to surgery.

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